Optimization of procedures for collecting and storing of CSF for studying the metabolome in ALS
Optimization of procedures for collecting and storing of CSF for studying the metabolome in ALS
复制标题
优化 CSF 收集和储存程序以研究 ALS 代谢组
DOI：
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发表时间：
2009
影响因子：
--
通讯作者：
P. Andersen
中科院分区：
文献类型：
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作者：
Anna Wuolikainen;M. Hedenström;T. Moritz;S. Marklund;H. Antti;P. Andersen
肌萎缩性侧索硬化症（ALS）的早期诊断、发展和治疗效果评估需要生物标志物。我们的目的是研究分析前因素是否会导致ALS患者脑脊液（CSF）代谢组学数据中的伪影。采用统计实验设计方案对16例患者的脑脊液进行研究，实验参数如下：储存温度（- 80°C/ - 20°C），收集管类型（聚丙烯/聚苯乙烯），收集到冷冻的时间延迟（0,10,30,90,150 min）。采用气相色谱-质谱法分析12例脑脊液，核磁共振谱法分析1例脑脊液。在另外3例患者中，研究了不同尺寸、不同温度、带/不带盖子的试管中采集的脑脊液中CO2的蒸发程度。我们发现，储存温度的变化对脑脊液代谢物组成的影响比其他任何研究的分析前参数都要大。CO2蒸发可能通过增加ph值而诱导代谢组中的伪影。总之，通过将CSF直接收集到N2(l)中密封严密的管中，然后将管冷冻至- 80°C以减少CO2蒸发，可以最大限度地减少所评估的伪影。
There is a need for biomarkers for early diagnosis, development and evaluation of treatment efficacy in amyotrophic lateral sclerosis (ALS). We aimed to investigate if pre-analytical factors induce artefacts in metabolomic data of cerebrospinal fluid (CSF) from patients with ALS. CSF from 16 patients was studied using a statistical experimental design protocol with the following parameters: storage temperature (−80°C/ − 20°C), type of collection tube (polypropylene/polystyrene), and time delay from collecting to freezing (0, 10, 30, 90, 150 min). Gas chromatography-mass spectrometry was used to analyse CSF from 12 of the patients while CSF from one patient was analysed with nuclear magnetic resonance spectroscopy. The extent of CO2 evaporization from CSF collected in tubes of different sizes at different temperatures and with/without lid were studied in three addtional patients. We found that alterations in storage temperature affect the metabolite composition of CSF more than any other studied pre-analytical parameter. CO2 evaporization may induce artefacts in the metabolome by increasing the pH. In conclusion, minimization of evaluated artefacts can be obtained by collecting the CSF directly into tubes with tightly sealed lids in N2(l) and after freezing transfer of the tubes to −80°C to minimize evaporation of CO2.
影响因子：
29
作者：
Chen, Chi;Shah, Yatrik M.;Gonzalez, Frank J.
通讯作者：
Gonzalez, Frank J.